{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,1]],"date-time":"2026-08-01T18:22:13Z","timestamp":1785608533703,"version":"3.56.0"},"reference-count":51,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,2,8]],"date-time":"2022-02-08T00:00:00Z","timestamp":1644278400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Flavivirus outbreaks require fast and reliable diagnostics that can be easily adapted to newly emerging and re-emerging flaviviruses. Due to the serological cross-reactivity among flavivirus antibodies, neutralization tests (NT) are considered the gold standard for sero-diagnostics. Here, we first established wild-type single-round infectious virus replicon particles (VRPs) by packaging a yellow fever virus (YFV) replicon expressing Gaussia luciferase (Gluc) with YFV structural proteins in trans using a double subgenomic Sindbis virus (SINV) replicon. The latter expressed the YFV envelope proteins prME via the first SINV subgenomic promoter and the capsid protein via a second subgenomic SINV promoter. VRPs were produced upon co-electroporation of replicon and packaging RNA. Introduction of single restriction enzyme sites in the packaging construct flanking the prME sequence easily allowed to exchange the prME moiety resulting in chimeric VRPs that have the surface proteins of other flaviviruses including dengue virus 1\u2013-4, Zika virus, West Nile virus, and tick-borne encephalitis virus. Besides comparing the YF-VRP based NT assay to a YF reporter virus NT assay, we analyzed the neutralization efficiencies of different human anti-flavivirus sera or a monoclonal antibody against all established VRPs. The assays were performed in a 96-well high-throughput format setting with Gluc as readout in comparison to classical plaque reduction NTs indicating that the VRP-based NT assays are suitable for high-throughput analyses of neutralizing flavivirus antibodies.<\/jats:p>","DOI":"10.3390\/v14020346","type":"journal-article","created":{"date-parts":[[2022,2,8]],"date-time":"2022-02-08T23:42:20Z","timestamp":1644363740000},"page":"346","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["High-Throughput Platform for Detection of Neutralizing Antibodies Using Flavivirus Reporter Replicon Particles"],"prefix":"10.3390","volume":"14","author":[{"given":"Arlen-Celina","family":"L\u00fccke","sequence":"first","affiliation":[{"name":"Institute of Virology, Medical Faculty, University of Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0814-5979","authenticated-orcid":false,"given":"Anja","family":"vom Hemdt","sequence":"additional","affiliation":[{"name":"Institute of Virology, Medical Faculty, University of Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janett","family":"Wieseler","sequence":"additional","affiliation":[{"name":"Institute of Virology, Medical Faculty, University of Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9163-2406","authenticated-orcid":false,"given":"Carlo","family":"Fischer","sequence":"additional","affiliation":[{"name":"Institute of Virology, Charit\u00e9-Universit\u00e4tsmedizin Berlin, Corporate Member of Freie Universt\u00e4t Berlin and Humboldt-Universit\u00e4t zu Berlin, 10117 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Feldmann","sequence":"additional","affiliation":[{"name":"Institute of Virology, Medical Faculty, University of Bonn, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1151-7614","authenticated-orcid":false,"given":"Simon","family":"Rothenfusser","sequence":"additional","affiliation":[{"name":"Division of Clinical Pharmacology, University Hospital, LMU Munich, 80337 Munich, Germany"},{"name":"Unit Clinical Pharmacology (EKliP), Helmholtz Center for Environmental Health, 80337 Munich, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan Felix","family":"Drexler","sequence":"additional","affiliation":[{"name":"Institute of Virology, Charit\u00e9-Universit\u00e4tsmedizin Berlin, Corporate Member of Freie Universt\u00e4t Berlin and Humboldt-Universit\u00e4t zu Berlin, 10117 Berlin, Germany"},{"name":"Martinovsky Institute of Medical Parasitology, Tropical and Vector-Borne Diseases, Sechenov University, 119435 Moskow, Russia"},{"name":"German Center for Infection Research (DZIF), Associated Partner Site Berlin, 10117 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9011-2764","authenticated-orcid":false,"given":"Beate Mareike","family":"K\u00fcmmerer","sequence":"additional","affiliation":[{"name":"Institute of Virology, Medical Faculty, University of Bonn, 53127 Bonn, Germany"},{"name":"German Center for Infection Research (DZIF), Associated Partner Site Bonn-Cologne, 53127 Bonn, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,8]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"796","DOI":"10.1038\/s41564-020-0714-0","article-title":"The continued threat of emerging flaviviruses","volume":"5","author":"Pierson","year":"2020","journal-title":"Nat. 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